Biotek Synergy HTX, Руководство оператора

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Fluorescence Tests |
127
•
Methylumbelliferone (“MUB”) (10-mg vial, BioTek PN 98156)
• Carbonate-Bicarbonate buffer (“CBB”) capsules (BioTek PN 98158)
• 100% methanol (BioTek PN 98161)
•
A new, clean, 96-well solid black plate, such as Corning Costar Mfr. #3915 or
equivalent.
• Excitation filter 360/40 nm installed
•
Emission filter 460/40 nm installed
•
Deionized or distilled water
• Various beakers, graduated cylinders, and pipettes
• Aluminum foil
•
(Optional, but recommended) 0.45 micron filter
•
Synergy HTX FI_MUB.prt
protocol described on page 129.
Test Solutions
1. Prepare the buffer (CBB) solution:
•
(Optional, but recommended) Using a 0.45-micron filter, filter 200 mL of
deionized or distilled water.
• Open and dissolve the contents of two CBB capsules (do not dissolve the
outer gelatin capsule) into 200 mL of the water.
• Stir the solution (preferably using a stir table) until the CBB is completely
dissolved.
2. Prepare the MUB stock solution:
• Add 1 mL of 100% methanol to the 10 mg vial of MUB.
• Make sure all of the dye has completely dissolved and is well mixed. This
yields a
10 mg/mL
stock solution.
• Wrap the solution in aluminum foil to prevent exposure to light.
3. Prepare the dilutions. Label each with “MUB” and the concentration.
Mix this MUB solution: with: to make:
0.5 mL of 10 mg/mL stock solution
4.5 mL of 100%
methanol
1 mg/mL
0.88 mL of 1 mg/mL solution 4.12 mL of CBB 176 μg/mL
0.1 mL of 176 μg/mL solution 9.9 mL of CBB 1.76 μg/mL
0.5 mL of 1.76 μg/mL solution 4.5 mL of CBB 176 ng/mL
1 mL of 176 ng/mL solution 9 mL of CBB
17.6 ng/mL
(100 nM)
Synergy HTX Operator's Manual